EP2254987B1 - Bioréacteur à perfusion pour fabrication et/ou culture d'un vaisseau sanguin humain ou animal et/ou d'un tissu humain ou animal - Google Patents

Bioréacteur à perfusion pour fabrication et/ou culture d'un vaisseau sanguin humain ou animal et/ou d'un tissu humain ou animal Download PDF

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EP2254987B1
EP2254987B1 EP09723875.2A EP09723875A EP2254987B1 EP 2254987 B1 EP2254987 B1 EP 2254987B1 EP 09723875 A EP09723875 A EP 09723875A EP 2254987 B1 EP2254987 B1 EP 2254987B1
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Prior art keywords
tissue
blood vessel
construct
bioreactor
chamber
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German (de)
English (en)
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EP2254987A2 (fr
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Bernard Frerich
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NOVATISSUE GmbH
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NOVATISSUE GmbH
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
    • A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
    • A61F2/02—Prostheses implantable into the body
    • A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
    • A61F2/06—Blood vessels
    • A61F2/062—Apparatus for the production of blood vessels made from natural tissue or with layers of living cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/22—Transparent or translucent parts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/26—Constructional details, e.g. recesses, hinges flexible
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/38—Caps; Covers; Plugs; Pouring means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/10—Perfusion
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/12—Pulsatile flow
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/14—Pressurized fluid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Definitions

  • the invention relates to a perfusable bioreactor for the production and / or cultivation of human or animal tissue or tissue equivalents, preferably in combination with a human or animal blood vessel or blood vessel equivalent.
  • constructs are artificially produced three-dimensional tissue equivalents which contain living cells in a three-dimensional matrix, in particular combinations of scaffolds and living cells, if appropriate also combined with matrix factors.
  • tissue equivalents and blood vessel wall equivalents are used analogously to this definition.
  • Investigations are the controlled provision of a relative overpressure in the vascular compartment - at least during the pulsation (systole) - against the "interstitium" as in a natural vessel and tissue. This is particularly difficult if a vessel wall equivalent is to be clamped, since its surface should remain observable at the same time.
  • the problem is to provide a seal that allows for at least a brief increase in pressure in the vascular compartment (through the pulse in a pulsatile perfusion).
  • a model that allows this in a small space can for many different issues in the fields of regeneration and tissue engineering, angiogenesis, circulatory and metabolic research, oncology (metastasis) u.v.a.m. be used.
  • the object of the invention is to provide a bioreactor which overcomes the disadvantages of the prior art.
  • the bioreactor has elastic walls in large proportions, has a tubular structure, and that a blood vessel, a blood vessel wall or a corresponding construct ("blood vessel equivalent") can be introduced in the interior such that it is arranged in the longitudinal axis of the bioreactor with a relative Overpressure relative to the ambient pressure (in bioreactor) can be perfused and also on transparent viewing window the possibility exists to observe it with optical magnification devices, eg laser scanning microscopes. All connections and openings for the introduction and manipulation of tissues, Scaffolds or scaffold cell combinations are introduced from the top of the head.
  • the overall diameter preferably does not exceed 17 mm, more preferably 13 mm, so that the bioreactor can be examined during operation in the well of benchtop MRI and ESR devices.
  • the perfusion takes place via a self-regulating perfusion system, which transports the perfusion medium and allows the setting of different perfusion modes (eg pulsation, frequency, etc.).
  • variant 1 the blood vessel wall or the blood vessel equivalent with its (endothelial) inside to the viewing window is arranged open.
  • the perfusable chamber is thus formed by the blood vessel wall / blood vessel wall equivalent, by the viewing window (which may also be made of elastic material) and by a circumferential rigid pressure surface.
  • the sealing of the chamber with respect to the remaining chamber volume is achieved by pressing the blood vessel wall / blood vessel wall equivalent with an elastic shaped body (eg elastic foam) against the circumferential pressure surface.
  • an elastic shaped body eg elastic foam
  • the pressure in the region of the circumferential pressure surface can be assisted by placing a second frame, which is congruent to the pressure surface, on the lower surface of the blood vessel wall / blood vessel wall equivalent, and furthermore that the elastic shaped body is constructed in a biphasic manner, ie with a less elastic peripheral edge supports the blood vessel wall / blood vessel wall equivalent in the area of the peripheral pressure surface and a higher elastic core, which promotes the pressure-induced deflections of the central area of the blood vessel wall / blood vessel wall equivalent.
  • a seal is provided circumferentially, which is not completely liquid-tight, but provides a seal to the extent that a short time a measurable pressure increase in the perfusable chamber occurs (up to the level of physiological and pathological Systolenhong ).
  • This basic construction offers the possibility of arbitrary between blood vessel wall / blood vessel wall equivalent and elastic shaped body Place tissue or tissue equivalent into which small blood vessels from the blood vessel wall / blood vessel wall equivalent einsprssen, which should be observed with the above imaging and measurement methods.
  • the entire remaining bioreactor cavity instead of the elastic shaped body to be filled with such a tissue or tissue equivalent, so that only the elastic bioreactor wall presses the blood vessel wall / blood vessel wall equivalent to the surrounding pressing surface and thus the sealing is achieved.
  • the perfusion medium may leave the perfusable chamber via the aforementioned drain and in addition through the tissue (perfusion ieS, eg, by grown capillaries or artificially created channels in the tissue piece) into the filled by foam / elastic molding material residual volume of the bioreactor, from where it has an additional Outflow exits the bioreactor.
  • a pulsatile perfusion via the tissue against the foam) or the elastic wall is passed and it can act mechanical or hydrodynamic stress on the tissue.
  • the essential difference to previous solutions lies in the fact that this perfusion dynamics takes place in an elastic environment and by adjusting the elasticity of the chamber wall, the compliance of natural blood vessels and tissue in physiological and pathological situation can be modeled.
  • the connections for monitoring systems can be integrated into the wall and / or the surrounding pressure surface.
  • the monitoring takes place via a probe system which monitors substance concentrations and physical or chemical parameters such as O 2 and CO 2 concentration, pressure in the chamber and in the remaining bioreactor, oxygen partial pressure, pH, flow rate and temperature , Elongation of the elastic walls can be monitored with strain gauges.
  • the monitoring also actively contributes to the regulation of growth conditions in the bioreactor system, since it is integrated into a control loop as sensor technology.
  • the perfusable chamber consists of a blood vessel or blood vessel equivalent which is guided along the viewing window and receives inflow and outflow through two ports.
  • the perfusable chamber can also be used with CLSM, MRI (magnetic resonance tomography) or ESR (electron spin resonance). Devices), but not by direct supervision of the endothelial surface.
  • CLSM magnetic resonance tomography
  • ESR electron spin resonance
  • bioreactor can be designed for single use.
  • the elastic sub-segment is more than 50% of the inner surface of the reactor wall 6, more preferably more than 75%. It is furthermore preferred that the elasticity of this material of the partial segment can be adjusted so that the expansion of tissue or tissue equivalent in the sheath by the perfusion pressure generated in the interior corresponds to the physiological or pathological values of the tissue compliance of the tissue to be produced. It is preferred that its overall diameter does not exceed 17 mm, more preferably 13 mm and in the well of ESR (electron spin resonance) devices and MRI (magnetic resonance imaging) devices can be examined.
  • ESR electron spin resonance
  • MRI magnetic resonance imaging
  • the head has at least on one side of the head a closable, preferably with screw cap 12 closable opening 4, via which the introduction of tissues, blood vessels, equivalents and moldings takes place. It is preferred that the perfusable pressure chamber 9 and viewing window 11 are dimensioned such that the vessel tissue piece 18 can be viewed with optical enlargers, especially fluorescence microscopes and confocal (laser scanning) microscopes.
  • a biphasic elastic material 20 which has a higher elasticity in its central portions 20.2 than in the edge region 20.1, so that the deflection of the vessel tissue piece is enhanced by the perfusion pressure in its central portion relative to the edge region and a support of the contact pressure of the lateral edge region of the construct is achieved.
  • the bioreactor contains internally immediately adjacent to and along the viewing window region a clamping device into which a blood vessel, a blood vessel equivalent or a framework for a tissue vessel to be manufactured by means of tissue engineering can be clamped between two tube connections 16.2 and 16.1 Blood vessel, the blood vessel equivalent or the framework for a tissue vessel to be manufactured by tissue engineering 21 through the viewing window from the outside and by means of optical devices, preferably confocal laser scanning microscopy, is observable.
  • the perfusable pressure chamber or blood vessel, blood vessel equivalent or scaffold for a tissue engineering engineering vessel 21 be perfused with culture medium or blood or a mixture of both. It is furthermore preferred that this be operable as a unit in conjunction with a self-regulating pulsatile perfusion system.
  • tissue can be exerted on the tissue via the perfusion dynamics, in particular by means of pulsatory perfusion with a self-controlling perfusion system. It is furthermore preferred that it contains resorbable or non-resorbable hollow fiber systems, conduit systems or framework structures in the interior, via which the culture medium is distributed, which in turn feed the tissue or tissue equivalent to be produced.
  • the bioreactor 1 is used for the production and / or cultivation of a human or animal blood vessel and / or a human or animal tissue (vascular tissue).
  • the bioreactor 1 has a tubular, hollow body 2, with two end faces 3.1 and 3.2.
  • On the front side 3.1 a resealable, liquid-tight opening for loading and assembly 4 is arranged, which serves in particular for entry of the construct 18 into the interior 5 of the reactor.
  • the opening 4 is formed as a conventional, liquid-tight screw cap 12.
  • a perfusable pressure chamber (perfusable chamber) 9 is arranged, which is at the same time a part of the inner space 5, which is arranged approximately parallel to the reactor longitudinal axis of the reactor 1 and which is open towards the reactor axis.
  • the inflow 7 opens into the pressure chamber (perfusable chamber) 9.
  • the pressure chamber (perfusable chamber) 9 has a circumferential pressure surface 8.1 the holder 8, to which liquid-tight the construct (tissue piece) 18, here a vessel wall equivalent is pressed.
  • the construct 18 thus forms a perfusable partition wall between the pressure chamber (perfusable chamber) 9 and the construct chamber 10, wherein the construct is thereby pressed against the pressing surface 8.1.
  • Part of the pressure chamber (perfusable chamber) 9 forms the monitoring window 11, which is designed as a transparent viewing window.
  • a construct chamber 10 which is arranged approximately parallel to the reactor longitudinal axis and is open towards the reactor axis or in the filled state, as in FIG. 1 represented by the construct is closed.
  • the outflow (discharge from the reactor chamber) 14 opens into the construct chamber 10 ..
  • the remaining cavity of the construct chamber 10, ie the space which is not filled in by the construct is, as in FIG. 1 represented by an elastic molded part (elastic molded body) 17 filled.
  • a part of the reactor wall 6, which is a part of the construct chamber 10, consists of elastic material, for example elastic silicone foil.
  • the tubular base body 2 is of the type dimensioned so that it can be inserted into the opening of an ESR or benchtop MRI device.
  • the front sides 3.1. and 3.2 are therefore each no larger than 13 mm in diameter (or 17mm for ESR).
  • in the FIG. 1 state shown is introduced into the bioreactor 1 via the inflow 7 a culture medium, blood, or the like. With pressure in the pressure chamber 9. A portion of this liquid medium flows through the perfusable construct and thus enters the region of the construct chamber 10. Via the outflow 14 and 15, liquid medium is led out of the bioreactor.
  • the liquid medium is thus supplied to a unit for generating pressure, for example a peristaltic pump.
  • a unit for generating pressure pulsating inflows can be generated, which then lie pulsating in the pressure chamber (perfusable chamber) 9.
  • This causes an at least partial deflection of the construct towards the construct chamber 10 and thus by displacing the contents of the construct chamber 10 also an expansion of the elastic reactor wall 6 outside.
  • Other arrangements are possible with the same basic principle ( Fig. 4 ).
  • a politicianssiedeltes flat vessel wall equivalent 18 for example consisting of a piece of pared with mesenchymal cells and endothelial cells to simulate a vessel wall to the size of the circumferential pressure surface 8.1 and placed flat on selbiger (endothelial side to the face plate 11 out).
  • a gap-shaped cavity 9 which corresponds to the perfusable chamber volume and can be viewed through the glass plate 11 directly from above is produced between glass plate 11 and tissue equivalent.
  • the side of the tissue facing the viewing plate 11 in this model corresponds to the inside of a blood vessel wall.
  • the viewing plate 11 allows the direct optical (eg by microscopy, fluorescence microscopy, laser scanning microscopy, etc.) monitoring of the developing surface of the vessel wall at the interface to the perfusion chamber 9 and also a non-invasive functional monitoring (eg by ESR spectroscopy, MRI) the vessel wall equivalent during reactor operation.
  • a tissue equivalent 19 for example, a scaffold with mesenchymal cells of a target tissue, eg muscle, fat tissue or bone is placed.
  • the remaining cavity of the bioreactor 1 is filled with an elastic foam.
  • the tissue equivalent / vessel wall equivalent is subjected, as needed, to physiological or pathological pressures and deflections measurable by pressure measurement systems inside the chamber.
  • a resilient fixation device (eg, a molded piece of foam) 17 formed in correspondence with the bioreactor interior is inserted into the bioreactor interior and presses the tissue piece 18 against the frame 8, thereby sealing the seal perfusable chamber volume 9 is achieved.
  • the foam thus fills the cavity between the tissue piece 18 and the flexible wall 6 and fixes the tissue piece 18 at the same time on the frame 8.
  • a solid or elastic plastic frame (additionally inserted frame) 8.3 in the extension of the frame on the face plate 11 between tissue piece 18 and foam 17 are placed so that the edge of the tissue piece 18 is fixed between two of the frame ( Fig. 3a ).
  • the diameter of the bioreactor interior can be reduced to fix the piece of fabric directly between the frame and the elastic wall, or make the piece of fabric or tissue equivalent 18 with a vessel wall or vessel wall equivalent 19 large enough to completely fill the cavity Fills without foam ( Fig. 3d Cross section through bioreactor 1 without shaped body, with vessel or vessel equivalent and attached tissue or tissue equivalent, completely filling the bioreactor space).
  • a biphasic elastic molded body / foam 20 can be used, in which the edge (solid zone) 20.1 is less elastic than the center (zone with high elasticity) 20.2. This ensures that the tissue piece in the center is more easily deflected by the pulsatory perfusion and better fixed and sealed at the edge ( Fig. 3b with biphasic molding and interposed frame and Fig. 3c with a biphasic molding without a frame set in two).
  • Example 4 Additional integration of a device for regulating the extensibility of the perfusable chamber
  • the faceplate can be modified as follows: For this, it is provided with a central, to which Size of the frame fitted recess provided and made a second support plate with the same recess. Now, a highly elastic membrane can be glued between the two support plates and acts from now on as a new section of the wall and as a regulator of compliance by expanding outward at each pulse and then tightened again. This membrane is preferably transparent so that the perfused tissue surface can be observed and microscoped through it.
  • the entire chamber wall is constructed of elastic material.
  • Example 5 Tubular vessel variant ( Figures 2a and b)
  • a variant embodiment is that there is no perfusable chamber ieS, but a slim holder 8 is integrated into the chamber wall and connects the two end faces, at the ends of each of which ports are attached, which serve to connect a blood vessel or blood vessel equivalent 21.
  • the connection to the inflow 7 must be designed so that the vessel / vessel equivalent 21 can be introduced in a sterile manner through the large opening 4 and coupled to the end face 3.2.
  • the face 3.2 is provided with a smaller opening with a flange, through which a coupling 16.2 can be introduced from the outside with a Schlaucholive on which the vessel / vessel equivalent 21 is fixed.
  • This coupling 16.2 for example, with liquid-tight Luer-Lock principle attached to the flange and fixes the vessel / vessel equivalent 21. Thereafter, the vessel / vessel equivalent 21 is attached to the port 16.1 (eg Schlaucholive).
  • the line 8.3 for the drain 15 is performed as in the first-described variant in the frame 8 or along it.
  • a tubular construct is perfused, all other features of the chamber are retained. Since the perfusion area can no longer be viewed, optical monitoring methods are more difficult than the first variant. In this way, however, a blood vessel can be produced or simulated by means of tissue engineering that is in direct contact with a supplied tissue section (construct, tissue piece) 18.
  • the conditions are examined, under which it comes to ingrowth of sprouts (small blood vessels) from the central vessel into the appended tissue (construct, tissue piece) 18.
  • sprouts small blood vessels
  • tissue construct, tissue piece
  • the monitoring window 11 is designed as a transparent film.
  • a self-regulating pulsatile perfusion system is connected to the bioreactor and serves to simulate physiological or experimental pressure conditions.
  • a comprehensive monitoring system connects to the frame's ports on the bioreactor's glass panel and allows real-time monitoring of critical parameters (O 2 , SpO 2 , CO 2 , pH, pressure, temperature, viscosity, flow rates, etc.).
  • critical parameters O 2 , SpO 2 , CO 2 , pH, pressure, temperature, viscosity, flow rates, etc.
  • bioreactor of the invention arise wherever the interactions between buttocks and stroma or mesenchymal or other Tissue play a role. These are many areas besides the already outlined applications in regenerative medicine and tissue engineering. As outlined in the previous examples, the system can be operated with natural, explanted tissues and vessels in the same way as tissue equivalents or artificial tissues. This results in a broad scope. This may be, for example, basic research, especially in the study of circulatory diseases, but also many metabolic disorders, such as obesity, in which the interplay of vessels and fat cells plays an essential role. Furthermore, it may be useful as a metastatic model in oncological research. Questions about wound healing can be answered, and it can also be used as an angiogenesis model in basic research. An essential branch is also the application in the testing of pharmaceuticals, eg the testing of the transfer of drugs into the interstitium or other questions. Here and in other applications, it can also be used as a substitute for animal testing.

Claims (15)

  1. Bioréacteur à perfusion pour la fabrication et/ou la culture d'un vaisseau sanguin humain ou animal et/ou d'un tissu humain ou animal, ce bioréacteur (1) comprenant au moins :
    - un corps principal (2) tubulaire, comportant deux faces terminales (3.1) et (3.2),
    - une ouverture (4) refermable étanche aux liquides, située sur une des faces terminales,
    - un espace interne (5),
    - une paroi de réacteur (6),
    - au moins une entrée (7) et au moins une sortie (14) pour un milieu liquide,
    - un dispositif de fixation (8) pour le tissu reconstitué à introduire dans le réacteur,
    caractérisé en ce que
    - dans l'espace interne se trouve une chambre de pression (9) à perfusion, qui est parallèle à l'axe longitudinal du réacteur,
    - au moins une entrée (7) et une sortie (15) débouchent dans la chambre de pression (9) et la chambre de pression (9) est ouverte sur l'axe du réacteur,
    - dans l'espace interne (5) se trouve une chambre de reconstitution tissulaire (10), qui est parallèle à l'axe longitudinal du réacteur et ouverte sur l'axe du réacteur,
    - au moins une sortie (14) débouche dans la chambre de reconstitution tissulaire (10),
    - le tissu reconstitué introduit dans le réacteur (1) peut être placé comme paroi de séparateur entre la chambre de pression (9) à perfusion et la chambre de reconstitution tissulaire (10) et
    - au moins un segment partiel de la paroi du réacteur (6), qui est
    au moins une partie de la chambre de reconstitution tissulaire (10), est composé d'une matière élastique.
  2. Bioréacteur d'après l'exigence 1, caractérisé en ce que dans la paroi du réacteur (6), en priorité dans le secteur de la chambre de pression (9), est située une fenêtre de contrôle (11) transparente, fixe ou élastique, ou la gaine du réacteur dans le secteur de la chambre de pression (9) ou le cas échéant également au-delà est fabriquée dans une matière transparente.
  3. Bioréacteur d'après l'exigence 1, caractérisé en ce que le bioréacteur peut être utilisé avec une surpression relative, la pression appliquée dans la chambre de pression (9) étant supérieure à celle dans la chambre de reconstitution tissulaire (10).
  4. Bioréacteur d'après l'exigence 1, caractérisé en ce que le dispositif de fixation (8) a au moins une surface de pression (8.1), le tissu reconstitué pouvant être pressé contre la surface de pression (8.1).
  5. Bioréacteur d'après l'exigence 1, caractérisé en ce que le dispositif de fixation (8) a au moins deux raccordements (16.1) et (16.2), entre lesquels un vaisseau sanguin tubulaire, l'équivalent de vaisseau sanguin ou une structure pour un vaisseau sanguin (21) à fabriquer au moyen de l'ingénierie tissulaire, peut être serré.
  6. Bioréacteur d'après l'exigence 1, caractérisé en ce que le régime charge de pression physique, qui est généré dans l'espace interne, correspond au régime charge de pression physique, qui influence en conditions normales physiologiques ou pathologiques dans l'organisme vivant humain ou animal sur les tissus, les équivalents de tissu, les vaisseaux sanguins ou les réseaux de vaisseaux sanguins, qui sont fabriqués.
  7. Bioréacteur d'après l'exigence 1, caractérisé en ce que toutes les entrées et les sorties ainsi que les raccordements pour les sondes aux faces terminales sont introduits dans le réacteur.
  8. Bioréacteur d'après l'exigence 1, caractérisé en ce qu'il dispose au moins sur un côté en haut, d'une ouverture (4) verrouillable, surtout en priorité verrouillable avec une fermeture à vis (12), qui permet l'introduction de tissus,
    de vaisseaux sanguins, d'équivalents et de corps moulés.
  9. Bioréacteur d'après l'exigence 1, caractérisé en ce que, entre le corps moulé élastique et le tissu reconstitué un deuxième cadre (8.3) coïncident au cadre (8) est introduit, ce deuxième cadre soutenant la pression de serrage du tissu reconstitué dans la zone du bord.
  10. Bioréacteur d'après l'exigence 1, caractérisé en ce que sur la face terminale (3.2) se trouve une deuxième petite ouverture, étanche aux liquides, par laquelle un vaisseau sanguin, un équivalent de vaisseau sanguin ou la structure pour un vaisseau sanguin à fabriquer au moyen de l'ingénierie tissulaire dans des conditions stériles peuvent être connectés à l'entrée (7), en particulier en priorité avec un raccord Luer-Lock modifié.
  11. Bioréacteur d'après l'exigence 1, caractérisé en ce qu'il ne nécessite aucun composant métallique.
  12. Procédé pour fabriquer des tissus, des équivalents de tissus, des vaisseaux sanguins ou des réseaux de vaisseaux sanguins humains ou animaux en utilisant un bioréacteur selon au moins une des exigences 1 à 11.
  13. Procédé pour fabriquer un système de vaisseaux sanguins artificiel, alimentant, fabriqué au moyen de l'ingénierie tissulaire en utilisant un bioréacteur selon au moins une des exigences 1 à 11, le procédé comprenant au moins les étapes suivantes : le tissu reconstitué est introduit dans l'espace interne du bioréacteur et l'espace creux restant de l'espace interne est rempli avec le milieu nutritif liquide et perfusé par l'entrée et la sortie avec un milieu nutritif.
  14. Utilisation d'un bioréacteur selon au moins une des exigences 1 à 11 pour la fabrication de tissus humains et animaux pour l'utilisation clinique et thérapeutique.
  15. Utilisation d'un bioréacteur selon au moins une des exigences 1 à 11 pour le test de substances pharmacologiques, en particulier dans le domaine de la recherche cardiovasculaire et de l'obésité, la cicatrisation des plaies et l'oncologie.
EP09723875.2A 2008-03-25 2009-03-23 Bioréacteur à perfusion pour fabrication et/ou culture d'un vaisseau sanguin humain ou animal et/ou d'un tissu humain ou animal Active EP2254987B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102008015633A DE102008015633B4 (de) 2008-03-25 2008-03-25 Perfundierbarer Bioreaktor zur Herstellung und/oder Kultivierung eines menschlichen oder tierischen Blutgefäßes und/oder eines menschlichen oder tierischen Gewebes
PCT/EP2009/002110 WO2009118141A2 (fr) 2008-03-25 2009-03-23 Bioréacteur à perfusion pour fabrication et/ou culture d'un vaisseau sanguin humain ou animal et/ou d'un tissu humain ou animal

Publications (2)

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EP2254987A2 EP2254987A2 (fr) 2010-12-01
EP2254987B1 true EP2254987B1 (fr) 2013-04-10

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US (1) US20110014597A1 (fr)
EP (1) EP2254987B1 (fr)
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WO (1) WO2009118141A2 (fr)

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US9456893B2 (en) 2011-07-29 2016-10-04 Southwest Research Institute Engineered tissue implants and methods of use thereof
DE102011082582B4 (de) * 2011-09-13 2015-06-25 Siemens Aktiengesellschaft Verfahren zur Überwachung eines Fermentationsprozesses
US8815594B2 (en) 2012-12-12 2014-08-26 Southwest Research Institute Hybrid tissue scaffold for tissue engineering
US9044530B2 (en) 2012-12-19 2015-06-02 Southwest Research Institute Fabrication of bone regeneration scaffolds and bone filler material using a perfusion flow system
WO2014174899A1 (fr) * 2013-04-23 2014-10-30 国立大学法人徳島大学 Cellule compressée ou tissu compressé en tant que modèle cellulaire d'affection pathologique et procédé de fabrication associé
DE102015210609B3 (de) * 2015-06-10 2016-01-14 OSPIN GmbH Modulares Bioreaktorsystem
ITUB20160272A1 (it) * 2016-01-22 2017-07-22 Univ Degli Studi Di Palermo Bioreattore a perfusione autosufficiente monouso per crescite cellulari 3D

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US5362622A (en) * 1993-03-11 1994-11-08 Board Of Regents, The University Of Texas System Combined perfusion and oxygenation apparatus
US6176874B1 (en) * 1993-10-18 2001-01-23 Masschusetts Institute Of Technology Vascularized tissue regeneration matrices formed by solid free form fabrication techniques
US5792603A (en) * 1995-04-27 1998-08-11 Advanced Tissue Sciences, Inc. Apparatus and method for sterilizing, seeding, culturing, storing, shipping and testing tissue, synthetic or native, vascular grafts
CA2370781A1 (fr) * 1999-04-30 2000-11-09 Joseph P. Vacanti Fabrication de tissus vascularises a l'aide de moules bidimensionnels microfabriques
DE19935643A1 (de) * 1999-07-29 2001-02-01 Augustinus Bader Vorrichtung zum Züchten und/oder Behandeln von Zellen
DE19964113A1 (de) * 1999-12-31 2001-07-05 Joerg C Gerlach Vorrichtung und Verfahren zur Züchtung und zur Nutzung von Hautzellen
DE10130512B4 (de) * 2001-06-25 2007-08-16 Bionethos Holding Gmbh Vorrichtung zur Druckperfusion für das Züchten und/oder für das Behandeln von Zellen
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DE10349484A1 (de) * 2003-10-21 2005-05-25 Universität Leipzig Verfahren und Bioreaktor zum Kultivieren und Stimulieren von dreidimensionalen, vitalen und mechanisch widerstandsfähigen Zelltransplantaten
DE102004012010A1 (de) * 2004-03-10 2005-09-29 Fachhochschule Gießen-Friedberg Erfindung betreffend Bioreaktoren und Bioreaktorsysteme
EP1693025A1 (fr) * 2005-02-17 2006-08-23 Universität Zürich Procédé pour la production d'une prothèse par génie tissulaire

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Publication number Publication date
DE102008015633A1 (de) 2009-10-01
EP2254987A2 (fr) 2010-12-01
WO2009118141A3 (fr) 2011-12-29
WO2009118141A2 (fr) 2009-10-01
DE102008015633B4 (de) 2010-07-01
US20110014597A1 (en) 2011-01-20

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